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First Record of American Lobster, Homarus Americanus (H. Milne Edwards, 1837), in the Mediterranean Sea

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First Record of American Lobster, Homarus Americanus (H. Milne Edwards, 1837), in the Mediterranean Sea BioInvasions Records (2020) Volume 9, Issue 1: 83–88 CORRECTED PROOF Rapid Communication First record of American lobster, Homarus americanus (H. Milne Edwards, 1837), in the Mediterranean Sea Mišo Pavičić*, Branko Dragičević, Iva Žužul, Dario Vrdoljak, Sanja Matić-Skoko and Tanja Šegvić-Bubić Institute of Oceanography and Fisheries, Šetalište I. Meštrovića 63, 21000 Split, Croatia *Corresponding author E-mail: [email protected] Citation: Pavičić M, Dragičević B, Žužul I, Vrdoljak D, Matić-Skoko S, Šegvić-Bubić Abstract T (2020) First record of American lobster, Homarus americanus (H. Milne Edwards, A live specimen of American lobster, Homarus americanus, was caught off Croatia, 1837), in the Mediterranean Sea. north Adriatic Sea. This is the first record of this species wild-caught in the BioInvasions Records 9(1): 83–88, Mediterranean. The origin of the caught specimen is unknown, yet possible pathways https://doi.org/10.3391/bir.2020.9.1.11 and potential impacts are discussed in the paper. Received: 28 June 2019 Accepted: 19 November 2019 Key words: introduced species, alien species, bioinvasion, Adriatic Sea Published: 12 January 2020 Handling editor: Christopher McKindsey Introduction Thematic editor: Cynthia McKenzie Copyright: © Pavičić et al. Occurrences of the non-native American lobster Homarus americanus, This is an open access article distributed under terms of the Creative Commons Attribution License recorded in European waters in recent decades, are likely related to live (Attribution 4.0 International - CC BY 4.0). seafood transport (D’Agaro et al. 2014; Øresland et al. 2017). Specimens of OPEN ACCESS. H. americanus have been occasionally captured in the waters of Sweden, Norway and Great Britain (Jørstad et al. 2007; Stebbing et al. 2012; Øresland et al. 2017). It has been imported live to many Mediterranean countries including Italy and Croatia (CABI 2013; SwAM 2016), a practice which represents a potential vector of introduction and, as a consequence, a potential threat to the economically important native European lobster, Homarus gammarus (Pavičić et al. 2018). Potential threats include competition (Øresland et al. 2017), transmission of diseases such as destructive epizootic shell disease (Stebbing et al. 2012), and possibility of hybridization (see in SwAM 2016). In this paper, we present the first record of wild-caught Homarus americanus in the Adriatic Sea, which also represents the first record of this species in Mediterranean waters. This finding is a matter of concern since there is evidence of its negative impact on native H. gammarus elsewhere (Øresland et al. 2017). Materials and methods In January of 2018, a live female American lobster was caught near the northwest coast of the Istrian peninsula in the north Adriatic Sea (Croatia; Pavičić et al. (2020), BioInvasions Records 9(1): 83–88, https://doi.org/10.3391/bir.2020.9.1.11 83 First record of American lobster in the Mediterranean Sea Figure 1. Location of the capture of Homarus americanus (indicated by the red dot). 45°30.502′N; 13°28.656′E) (Figure 1). The specimen was caught using a trammel net deployed at a depth of 20 m and transferred live to the Institute of Oceanography and Fisheries in Split, Croatia. The specimen was measured, weighed, and photographed. Morphological identification followed Holthuis (1991). A tissue sample from the 5th walking leg was stored in 96% ethanol for molecular species identification. DNA extraction, amplification and analysis Genomic DNA was isolated using a DNeasy 96 Tissue Kit (Qiagen) following the manufacturer’s instructions. After a DNA quality and quantity check, a fragment of the cytochrome oxidase subunit 1 gene (COI) was amplified by PCR reaction using a LCO and HCO primer pair (Folmer et al. 1994). PCR conditions were as follows: 15 min at 95 °C, 5 cycles of 94 °C for 30 s, 45 °C for 30 s and 72 °C for 1 min, followed by 35 cycles of 94 °C for 30 s, 50 °C for 30 s and 72 °C for 1 min, with final extension at 72 °C for 10 min. PCR products were visualized on 1% agarose gel under UV transilluminator. Product purification and sequencing were performed by Macrogen Inc. (Amsterdam, The Netherland) on an ABI 3730 automatic sequencer. For sequence identification, BLASTn (NCBI, available online) was used. Additionally, sequences of closely related species Pavičić et al. (2020), BioInvasions Records 9(1): 83–88, https://doi.org/10.3391/bir.2020.9.1.11 84 First record of American lobster in the Mediterranean Sea Figure 2. Female specimen of American lobster (Homarus americanus) caught in the Adriatic Sea. Photo by Mišo Pavičić. were obtained from GenBank for phylogenetic analyses. Sequence alignment was run by the ClustalW tool while phylogenetic analysis was carried out using Maximum likelihood (ML) approach in Mega v6 software (Tamura et al. 2013). The obtained COI sequence was deposited in GenBank (Accession Number: MN654341). Results and discussion The female specimen (Figure 2) measured 25.8 cm in total length (TL), 9.5 cm in carapace length (CL), and weighed 499 g. Main morphological traits which allowed identification of the specimen as H. americanus were as follows: presence of ventral spines on the rostrum (Figure 3A); brownish olive body coloration mottled with dark greenish black spots and pleura with reddish tips (Holthuis 1991; Jørstad et al. 2007). DNA barcoding was Pavičić et al. (2020), BioInvasions Records 9(1): 83–88, https://doi.org/10.3391/bir.2020.9.1.11 85 First record of American lobster in the Mediterranean Sea Figure 3. A) Spines on ventral side of rostrum (this specimen) of H. americanus; B) Absent spines on ventral side of rostrum of H. gammarus. Photos by Mišo Pavičić. Figure 4. Maximum-likelihood (ML) phylogenetic tree of the order Decapoda inferred from the cytochrome c coding (COI) region of the mtDNA. COI sequence data are labeled with its GenBank accession numbers. Numbers at nodes are bootstrap percentages after 1000 replicates based on distance. Scale bar represents the interval of the Kimura two-parameter (K2P) model. used to avoid possible misidentification and verify the identity of the specimen. Sequencing of the cytochrome oxidase subunit 1 fragment confirmed that the specimen is indeed the American lobster (H. americanus, Ass. No. MN654341). Namely, the 640 bp long mtDNA COI fragments of the studied specimen produced significant alignments with sequences of H. americanus from the Genbank with 99% similarity. Phylogenetic reconstruction (Figure 4) clearly aggregated our specimen with sequences of American lobster in a separate clade. The specimen is, to the best of our knowledge, the first record of H. americanus wild-caught in Mediterranean waters. We are unable to determine the origin and the vector of introduction of this specimen. However, we reason that its presence is likely due to live seafood importation to markets in the northern Adriatic Sea – the species has been Pavičić et al. (2020), BioInvasions Records 9(1): 83–88, https://doi.org/10.3391/bir.2020.9.1.11 86 First record of American lobster in the Mediterranean Sea imported live to Croatia by a large supermarket chain which distributes them to stores along the eastern Adriatic coast (pers. observation). Occurrences of introduced decapod crustaceans are already known from the Mediterranean Sea. For example, the North American blue crab, Callinectes sapidus, has profound impacts on certain habitats and ecosystems in the Mediterranean Sea and requires effective management measures (Perdikaris et al. 2016). Additionally, a single specimen of indo-pacific lobster, Panulirus ornatus (Galil 2007), was recorded in Israel in 1989. Its introduction was probably a consequence of migration through Suez Channel. However, its potential impact remains unknown. According to SwAM (2016), H. americanus and H. gammarus share similar habitat preferences. Beyond suggesting potential effects on native H. gammarus, this also provides support for the possibility of establishment of American lobster in the northern Adriatic, although H. gammarus is much less abundant in the area than it was historically, likely as a result of over- exploitation and global change (Phillips 2013). Since its mean of introduction is unknown and its status cannot be evaluated on the basis of a single record, any comment regarding its potential impact remains speculative. Taking into account European Parliament (2014) Regulation No 1143/2014, which urges the prevention and management of the introduction and spread of invasive alien species, in the future greater effort should be taken to establish better control of live animal trade, especially in cases when there is evidence that such animals could impact native species, as is the case with respect to H. gammarus. Acknowledgements We are thankful to Mr. Diego Makovac who provided the specimen. This work has been supported by Croatian Science Foundation (HRZZ) partially under the project LEKFishResCRO (IP-2016-06-5251) and partially under the project NurseFish (IP-2016-06-9884). We express our thanks to two anonymous reviewers for their helpful comments on the manuscript. References CABI (2013) Invasive Species Compendium. Datasheet Homarus americanus (American lobster), http://www.cabi.org/isc/datasheet/79674 (updated 6/11/2013) D’Agaro E, Sabbioni V, Messina MR, Tibaldi E, Bongiorno T, Tulli F, Lippe G, Fabbro A, Stecchini ML (2014) Effect of confinement and starvation on stress parameters in the American lobster (Homarus americanus). Italian Journal of Animal Science 13: 3530, https://doi.org/10.4081/ijas.2014.3530 European Parliament (2014) Regulation (EU) No 1143/2014 of the European Parliament and of the Council of 22 October 2014 on the prevention and management of the introduction and spread of invasive alien species. Official Journal of the European Union L317: 35–55 Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994) DNA primers for amplification of mitochondrial Cytochrome c Oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology 3: 294–299 Galil B (2007) Seeing Red: Alien species along the Mediterranean coast of Israel.
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